Determination of individual serum bile acids in chronic liver diseases: fasting levels and results of oral chenodeoxycholic acid tolerance test

Fifteen bile acids in serum of 5 normal subjects and 21 patients with chronic liver diseases were fractionated by high performance liquid chromatography. Fasting total bile acids (TBA), glycocholic acid, taurocholic acid, glycochenodeoxycholic acid (GCDCA), and taurochenodeoxycholic acid (TCDCA) were significantly increased in patients with liver cirrhosis as compared with normal subjects. The cholic acid (CA) level and the ratio of the sum of free and conjugated CA to the sum of free and conjugated chenodeoxycholic acid (CDCA) were significantly elevated in patients with compensated as compared with decompensated liver cirrhosis, and were useful for differentiation of the two conditions. Serum bile acid levels were determined after oral administration of 500 mg of CDCA in the 5 normal subjects and 11 patients with liver disease. The TBA level reached a peak 90 min after CDCA administration in patients with chronic hepatitis and after 120 min in those with liver cirrhosis. The increase in the TBA level was significantly greater in patients with liver disease than in normal subjects. CDCA, GCDCA, and TCDCA showed changes similar to those in TBA. In patients with decompensated liver cirrhosis, the reduction in the TBA and CDCA levels after the peaks was slow, and GCDCA and TCDCA levels continued to increase until 180 min after the administration of CDCA. The TBA and CDCA levels 180 min after CDCA administration were significantly different among normal subjects, patients with chronic hepatitis, those with compensated liver cirrhosis, and those with decompensated liver cirrhosis, suggesting the usefulness of CDCA administration in differentiation of these conditions.     

肝硬化患者中血清总胆汁酸测定的临床意义

背景：胆汁酸在肝内合成和分泌，因此可以作为反映肝细胞损害的指标之一。目的：探讨肝硬化患者血清总胆汁酸（TBA）测定的临床意义。方法：收集42例肝硬化患者的肝功能资料，比较TBA与其他常规肝功能指标的敏感性差异。结果：肝硬化组的TBA显著高于健康对照组（P＜0．01），其水平为健康对照组的6．9倍，异常率为74％，显著高于丙氨酸转氨酶（ALT）、γ－谷氨酰转移酶（γ－GT）和碱性磷酸酶（ALP）（P＜0．01）。肝硬化失代偿期患者的TBA水平显著高于代偿期患者（P＜0．01）。结论：TBA是反映肝硬化患者肝细胞损害的敏感指标之一。     

A combination therapy of vitamin K1 and bile acid on hemorrhagic diathesis in patients with decompensated liver cirrhosis

This paper presents a study of treatment involving vitamin K1 (VK1) accompanied by bile acids for hemorrhagic diathesis that was applied 42 times in 35 patients with decompensated liver cirrhosis. The hepaplastin test (HPT) value showed no change during the administration of VK1 alone. The HPT value elevated significantly, however, after the administration of VK1 with bile acid, especially when using ursodeoxycholic acid (UDCA). The HPT value in patients treated with VK1 in addition to UDCA before treatment with 53.2% +/- 10.2% (mean +/- SD) and after that for 2.1 +/- 1.1 months (mean +/- SD) with 74.7 +/- 16.8% showed a significant difference (p less than 0.001). On the other hand, no significant difference was noted between the HPT value of 57.2 +/- 13.6% before and that of 62.9 +/- 13.9% after the treatment in patients treated using VK1 in conjunction with chenodeoxycholic acid (CDCA). These results indicate that the therapy incorporating VK1 and bile acid, especially UDCA, is useful for reducing the hemorrhagic tendency in patients with decompensated liver cirrhosis who show no improvement using VK1 alone.     

胱抑素C、血浆尾加压素Ⅱ在失代偿期肝硬化合并肝肾综合征患者中的诊断价值

目的探讨胱抑素C（CysC）、血浆尾加压素Ⅱ（uⅡ）在失代偿期肝硬化合并肝肾综合征（HRS）患者中的诊断价值。方法收集2011年2月-2011年11月于常州市第一人民医院消化内科住院的失代偿期肝硬化患者50例,计算其校正内生肌酐清除率（Ccr）,并根据Ccr分为单纯肝硬化组、亚临床HRS组及HRS组,以15例健康体检者为对照组,检测ALT、AST、总胆汁酸（TBA）、Alb、血清肌酐（Scr）、CysC、uⅡ浓度,比较各组上述指标的差异,若数据成正态分布且方差齐,进行单因素方差分析（One-Way ANOVA）,两组间比较采用t检验;否则,多组间进行非参数秩和检验（Kruskall-Wallis H test）,两组间比较使用Mann-Whjtney U检验。相关分析采用Spearman相关检验。并通过绘制CysC、uⅡ浓度判断肝硬化合并亚临床HRS及HRS的ROC曲线,获得其曲线下面积（AUC）及最佳临界值。结果肝硬化病例组与正常对照组ALT、AST、TBA、Alb、Scr、Cysc、uⅡ比较差异有统计学意义。单纯肝硬化组、亚临床HRS组及HRS组CysC、uⅡ浓度均呈升高趋势,差异具有统计学意义（P〈0.05）。CysC和uⅡ判断亚临床HRS的AUC分别为0.91和0.867,其最佳临界值分别为1.40 mg/L、299.06 pg/ml。CysC和uⅡ判断HRS的AUC分别为0.942和0.901,其最佳临界值分别为2.22 mg/L、321pg/ml。结论 CysC、uⅡ浓度在患者血清肌酐正常时就出现升高改变,有助于早期发现亚临床HRS。CysC、uⅡ浓度对失代偿期肝硬化患者并发亚临床HRS及HRS有很好的判别价值。     

A combination therapy of vitamin K1 and bile acid on hemorrhagic diathesis in patients with decompensated liver cirrhosis

This paper presents a study of treatment involving vitamin K₁ (VK₁) accompanied by bile acids for hemorrhagic diathesis that was applied 42 times in 35 patients with decompensated liver cirrhosis. The hepaplastin test (HPT) value showed no change during the administration of VK₁ alone. The HPT value elevated significantly, however, after the administration of VK₁ with bile acid, expecially when using ursodeoxycholic acid (UDCA). The HPT value in patients treated with VK₁ in addition to UDCA before treatment with 53.2% ± 10.2% (mean ± SD) and after that for 2.1 ± 1.1 months (mean ± SD) with 74.7 ± 16.8% showed a significant difference (p< 0.001). On the other hand, no significant difference was noted between the HPT value of 57.2 ± 13.6% before and that of 62.9 ± 13.9% after the treatment in patients treated using VK₁ in conjunction with chenodeoxycholic acid (CDCA). These results indicate that the therapy incorporating VK₁ and bile acid, especially UDCA, is useful for reducing the hemorrhagic tendency in patients with decompensated liver cirrhosis who show no improvement using VK₁ alone.     

Assessment of the hepatic microvascular changes in liver cirrhosis by perfusion computed tomography

AIM： TO assess the hepatic microvascular parameters in patients with liver cirrhosis by perfusion computed tomography （CT）. METHODS： Perfusion CT was performed in 29 patients without liver disease （control subjects） and 39 patients with liver cirrhosis, including 22 patients with compensated cirrhosis and 17 patients with decompensated cirrhosis, proved by clinical and laboratory parameters. CT cine-scans were obtained over 50 s beginning with the injection of 50 mL of contrast agent. Hepatic microvascular parameters, mean transit time （MTT） and permeability surface area product （PS） were obtained with the Perfusion 3 software （General Electric, ADW 4.2）. RESULTS： The overall differences of MTT and PS between control subjects, patients with compensated cirrhosis and those with decompensated cirrhosis were statistically significant （P = 0.010 and P = 0.002, respectively）. MTT values were 15.613 ± 4.1746 s, 12.592 ± 4.7518 s, and 11.721 ± 4.5681 s for the three groups, respectively, while PS were 18.945 ± 7.2347 mL/min per 100 mL, 22.767 ± 8.3936 mL/min per 100 mL, and 28.735 ± 13.0654 mL/min per 100 mL. MTT in decompensated cirrhotic patients were significantly decreased compared to controls （P = 0.017）, whereas PS values were remarkably increased （P = 0.001）.CONCLUSION： The hepatic microvascular changes in patients with liver cirrhosis can be quantitatively assessed by perfusion CT. Hepatic microvascular parameters （MTr and PS）, as measured by perfusion CT, were significantly altered in decompensated cirrhosis.     

Total effective vascular compliance in patients with cirrhosis: a study of the response to acute blood volume expansion.

Although arterial vasodilation is a well-known feature in patients with cirrhosis, the venous system remains unexplored. To measure total effective vascular compliance, a reflection of the properties of the venous system, rapid volume expansion (300 ml of a gelatin solution in 3 min) was performed in 23 patients. Eleven patients had compensated cirrhosis (Child-Pugh grade A or B), and eight had decompensated cirrhosis (Child-Pugh grade C). Four control patients had mild chronic hepatitis, normal hepatic venous pressure and normal liver architecture. Cardiac index, hepatic venous pressures, hepatic and azygos blood flow and renal plasma flow were measured before and immediately after volume expansion. Right atrial pressure was recorded during volume expansion. This allowed the calculation of total effective vascular compliance, which was higher in patients with decompensated cirrhosis than in those with compensated cirrhosis (4.65 +/- 4.21 vs. 1.34 +/- 0.63 ml.mm Hg-1.kg-1; p less than 0.05). In response to volume expansion, renal vascular resistance decreased significantly in patients with compensated cirrhosis, but not in those with decompensated cirrhosis (-30% +/- 33% vs. +2% +/- 23%; p less than 0.05). No change was seen in glomerular filtration rate. Systemic oxygen consumption increased in patients with compensated cirrhosis, but not in those patients with decompensated cirrhosis (25% +/- 33% vs. -4% +/- 9%; p less than 0.05). Although in all patients with cirrhosis volume expansion increased central venous pressures, azygos blood flow and the hepatic venous pressure gradient did not change.(ABSTRACT TRUNCATED AT 250 WORDS)     

The relationship between postprandial bile acid concentration, GLP-1, PYY and ghrelin

Background Gut hormones peptide YY (PYY) and glucagon‐like peptide‐1 (GLP‐1) play an integral role in appetite control and energy homeostasis. Entero‐endocrine L‐cells can be stimulated by nutrients and or bile acids to co‐secrete PYY and GLP‐1. The aim of this study was to determine the response of bile acids, PYY, GLP‐1 and ghrelin after a test meal. Design Twelve subjects with a BMI of 22·8 (0·52) kg/m² [mean (SEM)] received a 400 kcal test meal after which blood samples were taken every 30 min from 0 to 180 min. PYY, GLP‐1 and ghrelin were measured by radioimmunoassays. Fractionated bile acids were measured by HPLC‐MSMS. Results PYY positively correlated with glycochenodeoxycholic acid (GCDCA) (rs = 0·23, P = 0·03) and taurochenodeoxycholic acid (TCDCA) (rs = 0·26, P = 0·02). GLP‐1 positively correlated with GCDCA (rs = 0·22, P = 0·047) and glycodeoxycholic acid (GDCA) (rs = 0·3, P = 0·005). Ghrelin negatively correlated with GDCA (rs = ?0·45, P≤ 0·0001), TCDCA (rs = ?0·23, P = 0·034) and taurodeoxycholic acid (TDCA) (rs = ?0·44, P≤ 0·0001). Conclusion PYY and GLP‐1 responses correlated with chenodeoxycholic acid (CDCA) counterparts, whereas ghrelin negatively correlated with deoxycholic acid (DCA) counterparts. Specific bile acids may thus differentially affect entero‐endocrine cells.     

乙型肝炎患者血脂、血清前白蛋白和总胆汁酸变化的意义

目的探讨乙型肝炎患者血脂、血清前白蛋白和总胆汁酸的变化规律,了解其与病情及预后的关系。方法在230例乙型肝炎患者[其中急性肝炎43例,慢性肝炎91例(轻度25例,中度32例,重度34例),重型肝炎33例,肝炎肝硬化63例]和50例正常人,测定血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL)、血清前白蛋白(PA)、白蛋白(ALB)、总胆汁酸(TBA)水平。结果各组总胆汁酸水平较正常对照组升高,差别有显著性意义(P<0·05);慢性肝炎组、肝硬化组、重型肝炎组患者血清甘油三脂、总胆固醇、HDL-C、血清前白蛋白、白蛋白水平均较正常对照组降低,差别有显著性意义(P<0.05);各项血脂水平随着肝功能损害的加重而降低(慢性肝炎重度组和中度组显著低于轻度组,重型肝炎组显著低于慢性肝炎组和肝硬化组)。结论检测乙型肝炎患者血脂、血清前白蛋白和总胆汁酸水平,对了解肝功能状态和估计预后均有一定的意义。     

Increase of sulfated ursodeoxycholic acid in the serum and urine of patients with chronic liver disease after ursodeoxycholic acid therapy

The present study was undertaken in order to investigate the influence of ursodeoxycholic acid (UDCA) on the composition of sulfate-conjugated bile acids in the serum and urine of patients with chronic active hepatitis and compensated liver cirrhosis. After a 12 week UDCA treatment (600 mg/day), total serum bile acid concentration increased two-fold in patients with compensated liver cirrhosis and increased slightly in patients with chronic active hepatitis. The percentage of sulfated bile acids significantly increased in patients with both compensated liver cirrhosis and chronic active hepatitis. UDCA made up 63% of the total serum bile acids in compensated liver cirrhosis and 61% in chronic active hepatitis after UDCA treatment. Of the serum bile acids after UDCA treatment, 35.2 and 53.9% of UDCA was sulfate conjugated in compensated liver cirrhosis and chronic active hepatitis, respectively. Urinary excretion of total bile acid and UDCA after UDCA treatment in compensated liver cirrhosis were higher than in chronic active hepatitis. UDCA made up 68% of the total urinary bile acids in compensated liver cirrhosis and 64% in chronic active hepatitis after UDCA treatment. Of the urinary bile acids after UDCA treatment, 51.8 and 54.8% of UDCA was sulfate conjugated in compensated liver cirrhosis and chronic active hepatitis, respectively. UDCA treatment for compensated liver cirrhosis was less effective than for chronic active hepatitis. We found that sulfate conjugation is one of the major metabolic pathways for UDCA after UDCA treatment in chronic liver diseases.     

Bile acids influence hepatic chemiluminescence in normal and oxidative-stressed rats†

The aim of the present study was to clarify whether bile acids influence chemiluminescence (CL) in the liver in vivo. Hepatic CL was determined on the surface of the liver of anaesthetized rats by using a photon counter. In normal rats, hepatic CL was significantly decreased 30 min after enteral administration of chenodeoxycholic acid (CDCA) or deoxycholic acid (DCA), but returned to its initial level 3 h later, after part of the CDCA administered was metabolized. Ursodeoxycholic acid (UDCA) and cholic acid had no effect on CL. In contrast, hepatic CL was markedly increased 30 min after CDCA or DCA administration in rats given either buthionine sulphoximine (BSO), an inhibitor of γ-glutamylcysteine synthetase, or diethyldithiocarbamate (DDC), an inhibitor of both superoxide dismutase and glutathione peroxidase. Chenodeoxycholic acid further increased the CL of BSO- or DDC-treated rats during inhalation of oxygen via a tracheal cannula. Coadministration of UDCA eliminated the effects of CDCA on the hepatic CL of normal and BSO- or DDC-treated rats with or without oxygen inhalation. We conclude that cytotoxic bile acids, such as CDCA, increase CL in the antioxidants-depleted or oxidative-stressed liver in vivc, but that UDCA prevents CDCA from developing CL.     

Feedback regulation of bile acid synthesis in primary human hepatocytes: evidence that CDCA is the strongest inhibitor

Primary human hepatocytes were used to elucidate the effect of individual bile acids on bile acid formation in human liver. Hepatocytes were treated with free as well as glycine-conjugated bile acids. Bile acid formation and messenger RNA (mRNA) levels of key enzymes and the nuclear receptor short heterodimer partner (SHP) were measured after 24 hours. Glycochenodeoxycholic acid (GCDCA; 100 micromol/L) significantly decreased formation of cholic acid (CA) to 44% +/- 4% of controls and glycodeoxycholic acid (GDCA) decreased formation of CA to 67% +/- 11% of controls. Glycoursodeoxycholic acid (GUDCA; 100 micromol/L) had no effect. GDCA or glycocholic acid (GCA) had no significant effect on chenodeoxycholic acid (CDCA) synthesis. Free bile acids had a similar effect as glycine-conjugated bile acids. Addition of GCDCA, GDCA, and GCA (100 micromol/L) markedly decreased cholesterol 7alpha-hydroxylase (CYP7A1) mRNA levels to 2% +/- 1%, 2% +/- 1%, and 29% +/- 11% of controls, respectively, whereas GUDCA had no effect. Addition of GDCA and GCDCA (100 micromol/L) significantly decreased sterol 12alpha-hydroxylase (CYP8B1) mRNA levels to 48% +/- 5% and 61% +/- 4% of controls, respectively, whereas GCA and GUDCA had no effect. Addition of GCDCA and GDCA (100 micromol/L) significantly decreased sterol 27-hydroxylase (CYP27A1) mRNA levels to 59% +/- 3% and 60% +/- 7% of controls, respectively, whereas GUDCA and GCA had no significant effect. Addition of both GCDCA and GDCA markedly increased the mRNA levels of SHP to 298% +/- 43% and 273% +/- 30% of controls, respectively. In conclusion, glycine-conjugated and free bile acids suppress bile acid synthesis and mRNA levels of CYP7A1 in the order CDCA > DCA > CA > UDCA. mRNA levels of CYP8B1 and CYP27A1 are suppressed to a much lower degree than CYP7A1.     

Urinary concentrations of bile acid glucuronides and sulfates in hepatobiliary diseases

Urinary bile acids in normal subjects and patients with obstructive jaundice and liver cirrhosis were quantitated by mass fragmentography after separation into nonglucuronidated-nonsulfated, glucuronidated and sulfated fractions. Mean values of total bile acids in urine were as follows: Control subjects (n = 7), 1.90 +/- 0.67; obstructive jaundice (n = 9), 77.90 +/- 40.39; liver cirrhosis, compensated (n = 6), 15.14 +/- 8.97, and decompensated (n = 6), 11.84 +/- 9.32 (mean +/- SD, mg/day). The percentages of each conjugate was 19-29% in the non-glucuronidated-nonsulfated fraction, 6-14% in the glucuronidated fraction and 60-74% in the sulfated fraction. Bile acids in urine and serum correlated well in each fraction (r = 0.82-0.84, p less than 0.001). The clearance of the three conjugates was the highest in the sulfates, and the clearance of glucuronides was higher than that of non-esterified bile acids. The glucuronidation and sulfation of bile acids play an important role in the detoxication of bile acids by excreting them into urine, especially in patients with elevated serum bile acids.     

酒精性肝硬化患者细胞因子改变的临床研究

目的探讨细胞因子在酒精性肝硬化发病机制中的作用。方法对酒精性肝硬化代偿期和失代偿期患者及健康者用放射免疫分析法检测细胞因子的含量进行分析。结果代偿期IL-6升高,差异有统计学意义（P〈0.01）,IL-10下降,差异有统计学意义（P〈0.05）;失代偿期IL-10无明显变化,差异无统计学意义（P〉0.05）,TGFβ、IL-6、IL-8、TNFα均升高,差异有统计学意义（P〈0.01）。结论酒精性肝硬化患者随着病情加重,细胞因子IL-6、IL-8、TNFα、TGFβ逐渐升高,IL-10无明显改变,对疾病的诊断及治疗有一定的指导作用。     

Changes in portal blood flow by intravenous administration of glucagon--the relationship between those changes and the total serum bile acid values in UDCA.GLUCAGON tolerance test

G30TBA is the total serum bile acid concentration obtained thirty minutes after injection of glucagon in the UDCA.GLUCAGON tolerance test. We have previously reported the usefulness of G30TBA in evaluating liver function. In this paper, changes in the portal blood flow in response to glucagon administration are measured and the relationship between those changes and G30TBA studied. We used an ultrasonic duplex system composed of a B-mode scanner and doppler flow meter of measure portal blood flow after injection of glucagon in thirty seven patients with chronic liver diseases. The results are as follows: The increase in portal blood flow by exogenous glucagon in liver cirrhosis is statistically significantly lower than that in chronic hepatitis. G30TBA in the good-response-to-glucagon group is statistically significantly lower than that in the poor-response group. These results suggest that the effect of glucagon on portal blood flow significantly influences the serum bile acid concentration in the UDCA.GLUCAGON tolerance test.     

Metabolism of 15N-ammonia in patients with cirrhosis: a three-compartmental analysis.

Urinary 15N-ammonia and 15N-urea were measured by gas chromatography-mass spectrometry after the intravenous administration of 15N-ammonia (0.2 mumol/kg/hr) to 6 volunteers and 11 patients with cirrhosis. Urinary 15N-nitrogen excretion as ammonia and urea was measured during the 210-min infusion period, and urea synthesis and ammonia conversion to amino acids were analyzed with a three-compartment model using the nonlinear least-squares method. The rate of urea synthesis in control subjects was 14.1 +/- 1.2 mg/kg/hr (mean +/- S.E.M.), and in cirrhotic patients it was 11.0 +/- 3.2 mg/kg/hr. The cirrhotic group was divided into those with compensated cirrhosis (Child class A patients) and those with decompensated cirrhosis (Child classes B and C patients), and the rates of urea synthesis for these groups were 14.5 +/- 1.5 and 8.9 +/- 1.6 mg/kg/hr, respectively. The difference between decompensated cirrhotic patients and control subjects was statistically significant (p less than 0.001). The percentage of ammonia reutilization of a given dose of 15N-ammonia was 75.9% +/- 2.4% in compensated cirrhotic patients and 82.9% +/- 3.6% in decompensated cirrhotic patients (p less than 0.05). Fasting venous ammonia levels correlated inversely with urea synthesis (p less than 0.001) and correlated positively with ammonia reutilization (p less than 0.05). These results are consistent with a decreased capacity to synthesize urea and an increased capacity to convert ammonia to amino acids in chronic liver failure.     

Clinical Significance of Fasting Serum Bile Acid in the Long-Term Observation of Chronic Liver Disease

Fasting serum bile acid (FSBA) was serially measured by a fluorescent enzyme method in a follow-up study of 61 patients with chronic liver disease. In chronic inactive hepatitis, fluctuation of FSBA was within the normal range in both the exacerbated state and in remission. In chronic active hepatitis, FSBA was abnormally elevated in both states, but the difference was not significant. In chronic active hepatitis where FSBA was elevated in the remission state above its value in the exacerbated state, exacerbation of the disease occurred repeatedly during the follow-up period. In compensated liver cirrhosis progressing into the decompensated form, FSBA levels increased before a decrease in the serum values of albumin, cholesterol, and cholinesterase, and an elevation of bilirubin. In liver cirrhosis, FSBA levels increased above 100 microM, 1-4 months before the appearance of ascites.     

Pharmacokinetics of Famotidine after Intravenous Administration in Liver Disease

The pharmacokinetics of famotidine were studied after the administration of a single intravenous dose of 20-mg to seven normal volunteers, six patients with chronic hepatitis, 14 patients with compensated cirrhosis, and seven patients with decompensated cirrhosis. The plasma terminal elimination half-life of famotidine was significantly prolonged and famotidine total body clearance was significantly reduced in patients with decompensated cirrhosis, whose creatinine clearance was 57.2 +/- 6.7 ml/min/1.48 m2, but these changes were not significant in patients with chronic hepatitis (creatinine clearance: 109.2 +/- 10.5 ml/min/1.48 m2) or in patients with compensated cirrhosis (creatinine clearance: 72.2 +/- 26.5 ml/min/1.48 m2 in comparison with normal volunteers. The total volume of distribution at steady state was not significantly different between the normal volunteers and the three groups of patients. Famotidine total body clearance showed a weak but significant correlation with the creatinine clearance (r = 0.66, p less than 0.001), serum albumin level (r = 0.51, p less than 0.01), and serum total bilirubin level (r = 0.36, p less than 0.05), which suggested that the reduction in clearance was due in part to the concomitant renal impairment, as well as hepatic dysfunction in these patients. In conclusion, famotidine total body clearance was reduced in decompensated cirrhosis, indicating that the dose schedule requires modification in patients with this condition.